ホ౯఍㆟ሗ࿌᭩
ER STTA ἲ㸦hERȘ-HeLa-9903 ⣽⬊ࢆ⏝࠸ࡓ࢚ࢫࢺࣟࢤࣥཷᐜయᜏᖖⓎ⌧⣔㌿෗άᛶ໬ヨ㦂ἲ㸧

JaCVAM ホ౯఍㆟

ᖹᡂ 28 ᖺ㸦2016 ᖺ㸧 12 ᭶ 6 ᪥11JaCVAM ホ౯఍㆟኱ 㔝 Ὀ 㞝 㸦㐠Ⴀጤဨ఍᥎⸀㸧㸸ᗙ㛗㣤 ሯ ᑦ ᩥ 㸦⊂❧⾜ᨻἲே ་⸆ရ་⒪ᶵჾ⥲ྜᶵᵓ㸧஬༑ᔒⰋ᫂㸦ᅜ❧་⸆ရ㣗ရ⾨⏕◊✲ᡤ㸧▼ ஭ 㞝 ஧ 㸦ᅜ❧་⸆ရ㣗ရ⾨⏕◊✲ᡤ Ᏻ඲ᛶ⏕≀ヨ㦂◊✲ࢭࣥࢱ࣮㸧ᒾ℩⿱⨾Ꮚ㸦᪥ᮏ〇⸆ᕤᴗ༠఍㸧㔠Ꮚ࿴ᘯ㸦᪥ᮏ໬Ꮫᕤᴗ༠఍㸧⠛ ⏣ ࿴ ಇ 㸦⊂❧⾜ᨻἲே ་⸆ရ་⒪ᶵჾ⥲ྜᶵᵓ㸧ᮡᒣ┿⌮Ꮚ㸦᪥ᮏ໬⢝ရᕤᴗ㐃ྜ఍㸧㇂ᕝᾈᏊ㸦᪥ᮏື≀ᐇ㦂௦᭰ἲᏛ఍㸧すᕝ⛅ె㸦ᅜ❧་⸆ရ㣗ရ⾨⏕◊✲ᡤ Ᏻ඲ᛶ⏕≀ヨ㦂◊✲ࢭࣥࢱ࣮㸧∾ ᰤ஧㸦᪥ᮏච␿ẘᛶᏛ఍㸧᳃⏣ ೺㸦᪥ᮏ⎔ቃኚ␗ཎᏛ఍㸧ᒣ⏣㝯ᚿ㸦⊂❧⾜ᨻἲே 〇ရホ౯ᢏ⾡ᇶ┙ᶵᵓ㸧ᶓ㛵༤㞝㸦᪥ᮏ⓶⭵࢔ࣞࣝࢠ࣮࣭᥋ゐ⓶⭵⅖Ꮫ఍㸧ྜྷ⏣Ṋ⨾㸦᪥ᮏẘᛶᏛ఍㸧ྜྷᮧ ຌ㸦ᗙ㛗᥎⸀㸧                ௵ᮇ㸸ᖹᡂ 26 ᖺ 4 ᭶ 1 ᪥㹼ᖹᡂ 28 ᖺ 3 ᭶ 31 ᪥኱ 㔝 Ὀ 㞝 㸦㐠Ⴀጤဨ఍᥎⸀㸧㸸ᗙ㛗㣤 ሯ ᑦ ᩥ 㸦⊂❧⾜ᨻἲே ་⸆ရ་⒪ᶵჾ⥲ྜᶵᵓ㸧஬༑ᔒⰋ᫂ 㸦ᅜ❧་⸆ရ㣗ရ⾨⏕◊✲ᡤ㸧▼ ஭ 㞝 ஧ 㸦ᅜ❧་⸆ရ㣗ရ⾨⏕◊✲ᡤ Ᏻ඲ᛶ⏕≀ヨ㦂◊✲ࢭࣥࢱ࣮㸧஭ ୖ ᬛ ᙲ 㸦᪥ᮏච␿ẘᛶᏛ఍㸧௒ ஭ ᩍ Ᏻ 㸦᪥ᮏື≀ᐇ㦂௦᭰ἲᏛ఍㸧ᒾ℩⿱⨾Ꮚ 㸦᪥ᮏ〇⸆ᕤᴗ༠఍㸧⠛ ⏣ ࿴ ಇ 㸦⊂❧⾜ᨻἲே ་⸆ရ་⒪ᶵჾ⥲ྜᶵᵓ㸧ᮡᒣ┿⌮Ꮚ 㸦᪥ᮏ໬⢝ရᕤᴗ㐃ྜ఍㸧௰ ஭ ಇ ྖ 㸦᪥ᮏ໬Ꮫᕤᴗ༠఍㸧୰ᮧࡿࡾࡇ 㸦⊂❧⾜ᨻἲே 〇ရホ౯ᢏ⾡ᇶ┙ᶵᵓ㸧す ᕝ ⛅ ె 㸦ᅜ❧་⸆ရ㣗ရ⾨⏕◊✲ᡤ Ᏻ඲ᛶ⏕≀ヨ㦂◊✲ࢭࣥࢱ࣮㸧἟ ⃝  ⪽ 㸦᪥ᮏẘᛶᏛ఍㸧᳃ ⏣  ೺ 㸦᪥ᮏ⎔ቃኚ␗ཎᏛ఍㸧ᶓ 㛵 ༤ 㞝 㸦᪥ᮏ⓶⭵࢔ࣞࣝࢠ࣮࣭᥋ゐ⓶⭵⅖Ꮫ఍㸧                ௵ᮇ㸸ᖹᡂ 28 ᖺ 4 ᭶ 1 ᪥㹼ᖹᡂ 30 ᖺ 3 ᭶ 31 ᪥22hERĮ-HeLa-9903 ⣽⬊ࢆ⏝࠸ࡓ࢚ࢫࢺࣟࢤࣥཷᐜయ㸦Estrogen receptor㸸ER㸧ᜏᖖⓎ⌧⣔㌿෗άᛶ໬ヨ㦂ἲ㸸The Stably Transfected TA assay using the human ERĮ-HeLa-9903 cell line㸦ERSTTA ἲࠊ௨ୗࠊᮏヨ㦂ἲ㸧ࡣࠊ໬Ꮫ≀㉁ࡢ࢚ࢫࢺࣟࢤࣥ㸦࠾ࡼࡧᢠ࢚ࢫࢺࣟࢤࣥ㸧άᛶࢆ
໬ᏛⓎග࡟ࡼࡾ᳨ฟࡍࡿ in vitro ヨ㦂ἲࡢ୍ࡘ࡛ࠊෆศἪ࠿ࡃ஘≀㉁ᑐ⟇ࡢࡓࡵ࡟㛤Ⓨࡉࢀࡓࡶࡢ࡛࠶ࡿࠋᮏヨ㦂ἲࡣ HeLa ⣽⬊࡟ᑟධࡉࢀࡓ ERĮ ࡢάᛶ໬࡟ࡼࡗ࡚㉳ࡇࡿ࣏࣮ࣞࢱ࣮㑇ఏᏊࡢ㌿෗άᛶࡢኚ໬ࢆ໬ᏛⓎග࡟ࡼࡾᐃ㔞ⓗ࡟ᐃࡍࡿヨ㦂⣔࡛࠶ࡿࠋᮏヨ㦂ἲࡢࣂࣜࢹ࣮ࢩࣙࣥ◊✲࡟ࡘ࠸࡚ࡣࠊERĮ ࢔ࢦࢽࢫࢺ࡜ ERĮ ࢔ࣥࢱࢦࢽࢫࢺ࡟ࡘ࠸࡚ࠊࡑࢀࡒࢀ⊂❧ࡋࡓ᪋タ㛫ࣂࣜࢹ࣮ࢩࣙࣥ◊✲ࡀᐇ᪋ࡉࢀࡓࠋ࢔ࢦࢽࢫࢺヨ㦂ࣂࣜࢹ࣮ࢩࣙࣥሗ࿌᭩ 1㸧ࡣࠊOECD㸦Organisation for Economic Co-operation and Development㸧࡟ 2006ᖺ࡟ᥦฟࡉࢀࠊ࢔ࣥࢱࢦࢽࢫࢺヨ㦂ࣂࣜࢹ࣮ࢩࣙࣥሗ࿌᭩ 2㸧ࡣࠊ2014 ᖺ࡟ᥦฟࡉࢀࡓࠋ๓⪅ࡣ 2009 ᖺ࡟ヨ㦂ἲ࢞࢖ࢻࣛ࢖ࣥ㸦Test Guideline: TG㸧455 ࡜ࡋ࡚ᢎㄆࡉࢀ 3)ࠊᚋ⪅ࡣ 2015
ᖺ࡟ TG455 ࡢᨵゞ∧࡜ࡋ࡚ TG ࡜࡞ࡗࡓ 4)ࠋJaCVAM㸦Japanese Center for Validation of
Alternative Methods㸧ホ౯఍㆟ࡣࠊෆศἪ࠿ࡃ஘ヨ㦂ἲ㈨ᩱ⦅⧩ጤဨ఍࡟ࡼࡾసᡂࡉࢀࡓࠕERSTTA ἲ㸸hERȘ-HeLa-9903 ⣽⬊ࢆ⏝࠸ࡓ࢚ࢫࢺࣟࢤࣥཷᐜయᜏᖖⓎ⌧⣔㌿෗άᛶ໬ヨ㦂ἲࡢホ౯ሗ࿌᭩ࠖ
㸦ᖹᡂ 28 ᖺ 10 ᭶ 21 ᪥㸧ࢆ⏝࠸࡚ࠊᮏヨ㦂ἲࡢጇᙜᛶ࡟ࡘ࠸᳨࡚ウࡋࡓࠋ㸯㸬ヨ㦂ἲࡢᐃ⩏ྡ⛠㸸 ER STTA ἲ㸦hERȘ-HeLa-9903 ⣽⬊ࢆ⏝࠸ࡓ࢚ࢫࢺࣟࢤࣥཷᐜయᜏᖖⓎ⌧⣔㌿෗άᛶ໬ヨ㦂ἲ㸧௦᭰ࡍࡿᑐ㇟ẘᛶヨ㦂㸸 in vivo ヨ㦂ἲࡢࠕࡆࡗṑ㢮ࢆ⏝࠸ࡿᏊᐑ⫧኱ヨ㦂ࠖ㸦OECD TG440,2007㸧5)ࠋࢆ௦᭰ࡍࡿヨ㦂ἲ࡛࠶ࡿࠋ㢮ఝࡢヨ㦂ἲ࡜ࡋ࡚ࠊin vitro ヨ㦂ἲࡢࠕࣛࢵࢺᏊᐑ࢚
ࢫࢺࣟࢤࣥཷᐜయ⤖ྜᛶヨ㦂ࠖ㸦OECD TG 493, 2015㸧6)࡜ࠕBG1Luc4E2 ⣽⬊ࢆ⏝࠸ࡿ࢚ࢫࢺࣟࢤࣥཷᐜయ㌿෗άᛶ໬ヨ㦂㸸BG1Luc estrogen receptor transactivation assay㸦BG1Luc ER TA㸧ࠖ㸦OECD TG 457, 2012㸧7)ࡀ࠶ࡿࠋ
ヨ㦂ἲࡢᴫ␎㸸 ⏕యࡢ ER ࡟⤖ྜࡋࠊ࢔ࢦࢽࢫࢺస⏝ࠊ࠶ࡿ࠸ࡣ࢔ࣥࢱࢦࢽࢫࢺస⏝ࢆ♧ࡍ໬Ꮫ≀㉁ࢆࢫࢡ࣮ࣜࢽࣥࢢࡍࡿࡓࡵ࡟ࠊᮏヨ㦂ἲࡣࠊHeLa ⣽⬊࡟ࣄࢺ ERȘࢆᜏᖖⓗ࡟Ⓨ⌧ࡍࡿࣉࣛࢫ࣑ࢻ࡜࢚ࢫࢺࣟࢤࣥᛂ⟅㓄ิ㸦Estrogen responsive element㸸ERE㸧ࡢୗὶ࡟ࣝࢩࣇ࢙࣮ࣛࢮ㑇ఏᏊࢆ⧅࠸ࡔ࣏࣮ࣞࢱ࣮ࣉࣛࢫ࣑ࢻࢆᑟධ࣭ᏳᐃⓎ⌧ࡉࡏࡓ⣽⬊㸦hERȘHeLa-9903 ⣽⬊㸧ࢆ⏝࠸ࡿࠋࡇࡢ⣽⬊࡟⿕㦂≀㉁ࢆ᭚㟢ࡋࡓᚋࡢࣝࢩࣇ࢙࣮ࣛࢮάᛶࡢኚ໬ࢆᐃࡍࡿࠋ㸰㸬ホ౯࡟⏝࠸ࡓ㈨ᩱ࠾ࡼࡧホ౯ෆᐜࡢ⛉Ꮫⓗጇᙜᛶ࢔ࢦࢽࢫࢺࢆホ౯ࡍࡿヨ㦂ἲ࡟㛵ࡋ࡚ࡣࠊ㸦୍㈈㸧໬Ꮫ≀㉁ホ౯◊✲ᶵᵓࡀ୰ᚰ࡜࡞ࡗ࡚ࠊ
4 ヨ㦂᪋タࡀཧຍࡋࡓࣂࣜࢹ࣮ࢩࣙࣥ◊✲ࡀᐇ᪋ࡉࢀࡓࠋࡑࡢሗ࿌᭩ࡀ 2006 ᖺ࡟ OECD ࡟
33ᥦฟࡉࢀࠊ2009 ᖺ࡟ TG455 ࡜ࡋ࡚ᢎㄆࡉࢀࡓࠋࡇࡢ TG455 ࡣࠊ2012 ᖺ࡟ BG1Luc ER TA ἲࡢ࢔ࢦࢽࢫࢺヨ㦂ἲࢆໟྵࡍࡿᛶ⬟‽ᣐヨ㦂ἲ࢞࢖ࢻࣛ࢖ࣥ㸦Performance-Based Test
Guideline㸸PBTG㸧8)࡟᭦᪂ࡉࢀࠊ࠶ࢃࡏ࡚࢔ࢦࢽࢫࢺヨ㦂ࡢᛶ⬟ᇶ‽㸦Performance Standards㸸PS㸧ࡀබ㛤ࡉࢀࡓ 9㸧ࠋ࢔ࣥࢱࢦࢽࢫࢺࢆホ౯ࡍࡿヨ㦂ἲ࡟㛵ࡋ࡚ࡣࠊJaCVAM ࡀ୰ᚰ࡜࡞ࡗ࡚ࠊ᭱⤊ⓗ࡟ 4 ヨ㦂᪋タ࡛ࣂࣜࢹ࣮ࢩࣙࣥ◊✲ࡀᐇ᪋ࡉࢀࡓࠋ࢔ࣥࢱࢦࢽࢫࢺࣂࣜࢹ࣮ࢩ
ࣙࣥሗ࿌᭩ࡣࠊඛ࡟ᥦฟࡉࢀࡓ࢔ࢦࢽࢫࢺ࡟ࡘ࠸࡚ࡢࣂࣜࢹ࣮ࢩࣙࣥሗ࿌᭩ࡢ㏣ຍᩥ᭩࡜ࡋ࡚ᥦฟࡉࢀࠊ᭱⤊ⓗ࡟࢔ࢦࢽࢫࢺホ౯ἲ㸦part A㸧࡜࢔ࣥࢱࢦࢽࢫࢺホ౯ἲ㸦part B㸧ࢆྵࡴ༢୍ࡢࣂࣜࢹ࣮ࢩࣙࣥሗ࿌᭩࡜ࡋ࡚බ㛤ࡉࢀࡓ 2㸧ࠋࡇࢀࡽࡢࣂࣜࢹ࣮ࢩࣙࣥሗ࿌᭩࡟ᇶ࡙
ࡁࠊ2014 ᖺ࡟࢔ࣥࢱࢦࢽࢫࢺヨ㦂ࢆໟྵࡍࡿ TG455 ᨵゞ᱌ࡀ OECD ࡟ᥦฟࡉࢀࠊᢎㄆࡉࢀ
ࡓࠋࡇࢀࡽࡢ㈨ᩱࢆ⏝࠸࡚ࠊJaCVAM ෆศἪ࠿ࡃ஘ヨ㦂ἲ㈨ᩱ⦅⧩ጤဨ఍ࡀホ౯ࡋࠊሗ࿌᭩
࡜ࡋ࡚ࡲ࡜ࡵࡓࡶࡢࢆホ౯㈨ᩱ࡜ࡋࡓࠋ
ᮏヨ㦂ἲࡣ HeLa ⣽⬊࡟ࣄࢺ ERȘࢆᑟධࡋࠊᙉไⓎ⌧ࡉࡏࡓ hER Ș-HeLa-9903 ⣽⬊ࢆ⏝࠸ࠊ࢔ࢦࢽࢫࢺ࠾ࡼࡧ࢔ࣥࢱࢦࢽࢫࢺࡢ୧⪅ࢆ᳨ฟ࡛ࡁࡿࡼ࠺࡞ in vitro ヨ㦂ἲ࡟ࡋࡓࡶ
ࡢ࡛࠶ࡿࠋỈࡸ Dimethyl sulfoxide㸦DMSO㸧➼࡟⁐ゎࡍࡿᗈ⠊࡞⿕㦂≀㉁࡟㐺⏝࡛ࡁࡿⅬ࡛౯್ࡀ㧗࠸ࠋ㢮ఝࡢ ER ㌿෗άᛶ໬ヨ㦂ἲ࡛࠶ࡿ BG1Luc ER TA ࡛⏝࠸ࡽࢀࡿ BG1Luc ⣽⬊ࡣࠊERȘ ᆺ㸭ERș ᆺ཮᪉ࢆⓎ⌧ࡋ࡚࠾ࡾࠊ୧⪅ࢆ௓ࡋࡓస⏝ࡀ᳨ฟྍ⬟࡛࠶ࡿ࡜ࡉࢀ࡚࠸ࡿࠋࡇࢀ࡟ᑐࡋ࡚ࠊᮏヨ㦂ἲ࡛ࡣࠊ⣧⢋࡟ࣄࢺ ERȘࡢࡳࢆ௓ࡋࡓస⏝ࢆ᳨ฟྍ⬟࡛࠶
ࡿࡀࠊ2 ✀ࡢ ER ࢔࢖ࢯࣇ࢛࣮࣒㸦ERȘࠊERș㸧࡟ᑐࡋ࡚᏶඲࡞㑅ᢥᛶࢆ♧ࡍ໬Ꮫ≀㉁ࡣ
▱ࡽࢀ࡚࠾ࡽࡎࠊᐃᛶⓗホ౯࡟࠾࠸࡚୧ヨ㦂ἲࡢ⤖ᯝࡣྠ➼࡜⪃࠼ࡽࢀࠊER ࢆ௓ࡋ࡚స⏝ࡍࡿ໬Ꮫ≀㉁ࢆࢫࢡ࣮ࣜࢽࣥࢢࡍࡿ᪉ἲ࡜ࡋ࡚⛉Ꮫⓗ࡞ጇᙜᛶࡀ࠶ࡿ࡜⪃࠼ࡽࢀࡿࠋ
㸱㸬ᮏヨ㦂ἲࡢ᭷⏝ᛶ࡜㐺⏝㝈⏺
ᮏヨ㦂ἲࡣ༢࡟࢔ࣥࢱࢦࢽࢫࢺ࡜ ER ࡜ࡢ⤖ྜ࡟࡜࡝ࡲࡽࡎ࢔ࢦࢽࢫࢺࡢάᛶࢆᢚไࡍ
ࡿຠᯝࢆ᳨ฟ࡛ࡁࡿⅬࡀඃࢀ࡚࠸ࡿࠋICCVAM㸦Interagency Coordinating Committee on theValidation of Alternative Methods㸧ཧ↷ศ㢮ࠊER ⤖ྜヨ㦂ࠊᏊᐑ⫧኱ヨ㦂➼ࡢ⤖ᯝ࡜ࡢ୍⮴ᗘࠊᮏヨ㦂ἲࡢឤᗘࠊ≉␗ᗘࡣⰋዲ࡛࠶ࡾࠊഇ㝧ᛶࡸഇ㝜ᛶࡣᑡ࡞࠸࡜࠸࠺⤖ᯝࢆ♧ࡋ࡚࠸ࡿࠋ
BG1Luc ER TA ヨ㦂ἲ࡟ẚࡋឤᗘࡀ㧗ࡃࠊ࢔ࢦࢽࢫࢺࠊ࢔ࣥࢱࢦࢽࢫࢺࢆ 1/100 ࡢ⃰ᗘ᳨࡛ฟ࡛ࡁࡿࠋᮏヨ㦂ἲ࡟ࡼࡿ໬Ꮫ≀㉁ࡢ࢔ࢦࢽࢫࢺάᛶ࣭࢔ࣥࢱࢦࢽࢫࢺάᛶࡢࢫࢡ࣮ࣜࢽࣥࢢࡢ⤖ᯝࢆ ICCVAM ཧ↷ศ㢮࡜ࡢẚ㍑ࡋࡓ⤖ᯝࠊ୍⮴⋡ࡣ 100㸣࡛࠶ࡗࡓࠋ࢚ࢫࢺࣟࢤࣥάᛶࡢ᳨ฟ࡟⏝࠸ࡽࢀࡿ᪤Ꮡࡢ⣽⬊⣔㸦ෆᅾᛶࡢࣄࢺ ER ࢆ฼⏝ࡍࡿ BG1 ⣽⬊㸧࡞࡝࡜ẚ㍑ࡋ࡚
ࡶࠊ㝧ᛶ≀㉁ࡸ㝜ᛶ≀㉁ࡢ㆑ูᛶࡣⰋዲ࡛࠶ࡿࠋᮏヨ㦂ἲ࡛⏝࠸࡚࠸ࡿ hERȘ-HeLa-9903⣽⬊ࡣቑṪࡀ᪩࠸ࡓࡵࠊヨ㦂‽ഛࡢࡓࡵࡢ᫬㛫ࢆ▷ࡃࡍࡿࡇ࡜ࡀ࡛ࡁࠊከᩘࡢ໬Ꮫ≀㉁ࡢࢫ
ࢡ࣮ࣜࢽࣥࢢ࡟ྥ࠸࡚࠸ࡿࠋ࣏࣮ࣞࢱ࣮࡜ࡋ࡚⏝࠸࡚࠸ࡿࣝࢩࣇ࢙࣮ࣛࢮάᛶ࡟ᙳ㡪ࢆ୚࠼ࡿ໬Ꮫ≀㉁࡛ࡣࠊER 㠀≉44␗ⓗ࡞ஹ㐍ࡸ㜼ᐖ➼࡟ࡼࡾഇ㝧ᛶ㸦ࡶࡋࡃࡣഇ㝜ᛶ㸧཯ᛂࢆច㉳ࡍࡿྍ⬟ᛶࡀ࠶ࡿࡓࡵὀពࡀᚲせ࡛࠶ࡿࠋER ࢆ௓ࡋࡓ໬Ꮫ≀㉁ࡢ࢚ࢫࢺࣟࢤࣥάᛶࢆ୍ḟࢫࢡ࣮ࣜࢽࣥࢢࡍࡿ࡟ࡣ౽฼࡞ヨ㦂⣔࡛࠶ࡿࡀࠊ⣽⬊ࡢ⥅௦࡟ࡼࡗ࡚ ER ࡢ཯ᛂᛶࡀኚ໬ࡋ࡞࠸ࡇ࡜ࢆ☜ㄆࡋ࡚࠾ࡃᚲ
せࡀ࠶ࡿࠋOECD TG ࡛♧ࡉࢀࡓ ERȘ࢔ࢦࢽࢫࢺ࠾ࡼࡧ ERȘ࢔ࣥࢱࢦࢽࢫࢺヨ㦂ࡑࢀࡒࢀࡢ⇍㐩ᗘ☜ㄆ≀㉁ 14 ≀㉁࡜ 10 ≀㉁ࢆ⏝࠸࡚⏝㔞-ᛂ⟅ᛶࢆㄪ࡭࡚࠾ࡃࡇ࡜ࡀᚲせ࡛࠶ࡿࠋࡲࡓࠊ⁐፹࡜ࡋ࡚Ỉࡸ Ethanol (>95%)࠶ࡿ࠸ࡣ DMSO ࡀ⏝࠸ࡽࢀ࡚࠸ࡿࡀࠊDMSO ࡟⁐ࡅ࡟ࡃ࠸≀㉁࡟ࡘ࠸࡚ࡣࠊ௚ࡢ᭷⏝࡞⁐፹ࢆ᳨ウࡋࠊࡑࢀࡀ hERȘ-HeLa-9903 ⣽⬊࡟ᙳ㡪ࡋ࡞࠸ࡇ࡜ࢆ᳨ドࡍࡿᚲせࡀ࠶ࡿࠋ⌧᫬Ⅼ࡛ࡣ᥹Ⓨᛶ≀㉁ࡢྲྀࡾᢅ࠸࡟ࡘ࠸࡚᫂☜࡞ᣦ㔪ࡀ↓࠸ࠋ௒ᚋࡢ᳨ウࡀᮇᚅࡉࢀࡿࠋࡲࡓࠊ௦ㅰࡉࢀ࡚࠿ࡽ ERȘ࢔ࢦࢽࢫࢺ࠾ࡼࡧ ERȘ࢔ࣥࢱࢦࢽࢫࢺస⏝ࢆ♧ࡍ≀㉁ࡢホ౯ࡶ⾜࠼࡞࠸ࠋ㸲㸬┠ⓗ࡜ࡍࡿ≀㉁ཪࡣ〇ရࡢẘᛶࢆホ౯ࡍࡿヨ㦂ἲ࡜ࡋ࡚ࡢࠊ♫఍ⓗཷࡅධࢀᛶ࠾ࡼࡧ⾜ᨻୖࡢ฼⏝ࡢྍ⬟ᛶ ♫఍ⓗཷࡅධࢀᛶ㸸ᮏヨ㦂ἲࡣ㑇ఏᏊ⤌ࡳ᥮࠼࡟ࡼࡾస〇ࡉࢀࡓ hERȘ-HeLa-9903 ⣽⬊ࢆ⏝࠸ࡿヨ㦂ἲ࡛࠶ࡾࠊ⏕ࡁࡓື≀ࢆ⏝࠸࡞࠸࡜࠸࠺Ⅼ࡛ࠊ3Rs ࡢ⢭⚄࡟ྜ⮴ࡋ࡚࠸ࡿࠋࡇࡢヨ㦂࡛ᚲせ࡞ᢏ⾡ࡣࠊᇵ㣴⣽⬊ࢆ⏝࠸ࡿヨ㦂ἲ୍⯡ࡢᢏ⾡࠾ࡼࡧ⣽⬊ࡢⓎගࢆᐃࡍࡿᢏ⾡࡛࠶ࡾࠊ㐺ษ࡞カ⦎࡟ࡼࡗ࡚ᐜ᫆࡟⩦ᚓ࡛ࡁࡿࡶࡢ࡛࠶ࡿࠋࡲࡓࠊᮏヨ㦂ࡢࡓࡵ࡟ᚲせ࡞ᶵჾࡣࠊ㏻ᖖࡢ⣽⬊ᇵ㣴࡟せࡍࡿ⿦⨨ࡢ࡯࠿ࠊ⣽⬊Ⓨගࡢᐃ࡟⏝࠸ࡿගᗘィ࡛࠶ࡾࠊ㧗౯࡞ࡶࡢ࡛࡞࠸ࠋ⣽⬊ࡶබⓗ࡞⣽⬊ࣂࣥࢡ࠿ࡽධᡭྍ⬟࡛࠶ࡿࠋ௨ୖࡼࡾࠊᮏヨ㦂ἲࡢ♫఍ⓗཷࡅධࢀᛶࡣ㧗࠸࡜⪃࠼ࡿࠋ ⾜ᨻୖࡢ฼⏝ᛶ㸸 ᮏヨ㦂ἲࡣᇵ㣴⣽⬊ࢆ⏝࠸ࡿ in vitro ヨ㦂ἲ࡛࠶ࡾࠊ໬Ꮫ≀㉁ࡢ࢚ࢫࢺࣟࢤࣥཷᐜయ࡬ࡢ
స⏝ࡢ᭷↓ࢆホ౯࡛ࡁࠊㄗホ౯ࡀᑡ࡞࠸ࡇ࡜࠿ࡽࠊ㢮ఝヨ㦂ἲ࡛࠶ࡿ BG1Luc ER TA ἲ࡜ྠࡌ⛬ᗘ࡟ 10㸧ࠊ⾜ᨻୖ฼⏝ࡀྍ⬟࡛࠶ࡿ࡜⪃࠼ࡿࠋᘬ⏝ᩥ⊩1) Takeyoshi, M. (2006), Draft Report of Pre-validation and Inter-Laboratory Validation for Stably
Transfected Transcriptional Activation (TA) Assay to Detect Estrogenic Activity - The Human
Estrogen Receptor Alpha Mediated Reporter Gene Assay Using hER-HeLa-9903 Cell Line,
Chemicals Evaluation and Research Institute (CERI): Japan. p1-188.
2) OECD (2015), Report of the inter-laboratory validation for stably transfected transactivation assayto detect estrogenic and anti-estrogenic activity, Series on Testing & Assessment No. 225.
3) OECD (2009), Test No. 455: The Stably Transfected Human Estrogen Receptor-alpha55

Transcriptional Activation Assay for Detection of Estrogenic Agonist-Activity of Chemicals
4) OECD (2015), Test No. 455: Performance-Based Test Guideline for Stably TransfectedTransactivation In Vitro Assays to Detect Estrogen Receptor Agonists and Antagonists5) OECD (2007), Test No. 440: Uterotrophic Bioassay in Rodents
6) OECD (2015), Test No. 493: Performance-Based Test Guideline for Human RecombinantEstrogen Receptor (hrER) In Vitro Assays to Detect Chemicals with ER Binding Affinity
7) OECD (2012) Test No. 457: BG1Luc Estrogen Receptor Transactivation Test Method for
Identifying Estrogen Receptor Agonists and Antagonists8) OECD (2012), Test No. 455, Performance-Based Test Guideline for Stably TransfectedTransactivation In Vitro Assays to Detect Estrogen Receptor Agonists and Antagonists
9) OECD (2012) Performance standards for stably transfected transactivation in vitro assays to detect
estrogen agonists for TG 455, Series on Testing & Assessment No. 173.
10) JaCVAM ホ౯఍㆟ሗ࿌᭩㸦ᖹᡂ 25 ᖺ 6 ᭶ 11 ᪥㸧ࣄࢺ࢚ࢫࢺࣟࢤࣥཷᐜయ⤖ྜ࡟ࡼࡿάᛶ໬࣭ᣕᢠస⏝≀㉁ࢆ᳨ฟࡍࡿ BG1Luc ER TA ἲ66